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Two-dimensional immunoelectrophoresis was used to study the antigenic composition and serological relationships of the following five Acholeplasma species: Acholeplasma axanthum, Acholeplasma eqir$etale, Ac-holeplusma grun- irlarum, Acholeplasma hippikon, and Acholeplasmu laidlmii. A total of 15 to 25 precipitin peaks were observed in the homologous reactions. A. equijetale and A. hippikon were most closely related serologically, having six related antigens. Nearly all antigens were negatively charged at pH 8.6: the only exceptions were one antigen of A. uxanthum and one antigen of A. laidlawii. The "positively charged" antigens were membrane associated and did not cross-react, One common antigen was recognized in each organism by all heterologous antisera. Monospecific antiserum prepared against the common antigen (peak 0.72 of A. hippikon) reacted with one antigen in each species tested. The electrophoretic mobilities of the common antigen ranged from 0.45 (A. axanthrrm) to 0.72 (A. hippikon). This antigen was located in the cytoplasm of A. laidlntrlii because the antiserum reacted only with the cytoplasmic fractions, not with the membrane fractions. The recognition of a common cytoplasmic antigen in the Acholeplasma- taceae is important to the taxonomy of these organisms. The organisms classified in the genus Achole- plasma are presently separated into a number of species, including Acholeplasmu axanthum, Acholeplasma equifetale, Acholeplasma granir- larum, Acholeplasma hippikon, Acholeplusma 1 a idlu w ii , A c h o 1 ep la s m u rn odic u m , A c'h ol e - plasma morum, and Acholeplasma oculi, on the basis of serological distinctions and a limited number of biochemical and physiological differ- ences (18). Double-immunodiffusion studies of the Acholeplasma species have shown that these organisms have one or more common antigens (8, 9). However, the double-immunodiffusion technique is relatively insensitive, and neither antigens nor antibodies can be quantitated (8). In this study, we used the two-dimensional immunoelectrophoretic method of Laurel1 (11) to study the antigenic composition and serolog- ical relationships of five of the seven Achok- plasma species because of its great sensitivity and resolving power. A monospecific antiserum against one antigen common to the genus was prepared and was used to identify the common antigen in the various species.

Common Cytoplasmic Antigen in Five Acholeplasma Species